EV Profiler Kit

Quantify EVs with nanometer-precision

Harness the power of super-resolution microscopy to characterize EV populations and individual EVs. Quantify biomarkers on small, rare, and precious EVs with single molecule sensitivity. ONI’s EV Profiler helps you focus on answering questions about your EVs.

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Identify EVs and quantify surface biomarkers

With the EV Profiler Kit you use our validated antibodies to measure the expression of tetraspanins and characterize the single, double and triple positive populations, or you can measure your own biomarker of interest.

Flexible Capture of your EVs

Capture with phosphatidyl-serine-based or antibody-based capture.

Assess EV morphology

Single molecule localization microscopy used in combination with the appropriate fluorescently labeled probes, helps assess the integrity and morphology of EV populations and single EVs after purification. ONI’s EV Profiler Kit in combination with cluster-based analysis on our CODI cloud platform, allows reporting of morphological features of the EV, including size and circularity.

Detection of EV cargo and surface biomarkers on the same sample

The EV Profiler Kit V2 is able to detect and quantify luminal protein cargo on single EVs, as well as surface biomarkers. Up to 3 markers can be detected within the same sample.

The EV Profiler Kit V2 is able to detect and quantify luminal protein cargo on single EVs, as well as surface biomarkers. HCT116-derived EVs were stained with ONI’s pan-EV stain labeled with 488, Tetraspanin Trio (anti-CD9, anti-CD63 and anti-CD81 all conjugated to 568 and anti-Syntenin-1 labeled with 647. Graphs show the number of Tetraspanins or Syntenin-1 counts from EVs positive for either or both markers. Permeabilization increases the overlap of Tetraspanin and Syntenin-1 counts from 0.61% to 35.8% (percentages not shown). 

Characterize across-scales: from EV populations down to single EVs

Study EV populations across scales. Each EV Profiler kit contains enough material to analyze 12 EV samples using our 4 lane analysis chip which has been developed and optimized for EV capture. You can capture and image hundreds of EVs from any given source with the appropriate capture method (read more details below), then zoom into individual EVs to see biomarker distribution at a single-EV level.

ONI’s Nanoimager and CODI platform enable imaging and visual inspection of EV samples across scales. Scale bars range from 10 µm to 50 nm when inspecting one EV out of thousands of EVs imaged on from an HCT116-derived EV sample captured with ONI’s S4 molecule and labeled with pan-EV stain (yellow), anti-CD63-568 (cyan) and anti-CD81-647 (magenta).

Obtain rapid results with a benchtop assay

Capture, stain, image and quantify

EV Profiler V2 contains all necessary reagents to capture, fix and stain EVs ready for imaging in a same-day workflow for single EV characterization. The workflow works with EV sample volumes as low as 10 µL.

Single molecule localization data and nanoscale resolution images can be obtained using the Nanoimager, our benchtop super-resolution microscope. Data is then analyzed through our CODI cloud-based software, making it possible to get results within a few hours from start to finish. 

Flexibility to suit your EV samples

Multiple capture strategies

EV Profiler offers 3 options for EV capture on the passivated glass surface of the assay chips:

Tetraspanin based capture

ONI validated capture antibodies against tetraspanins: anti-CD9, anti-CD63 and anti-CD81 antibodies are included in the kit

Phosphatidylserine-dependent, protein-independent capture

Our proprietary S4 capture molecule binds EVs in a protein-independent manner, working best with phosphatidylserine-rich EVs.

Custom target based capture

Use your own antibody of choice against your target of interest – it simply needs to be biotinylated (sample above provided by Dr Nicole Meisner-Kober’s lab).

Swap our antibodies for your own easily

In the EV Profiler Kit V2 antibodies for standard tetraspanin markers (CD9, CD63, and CD81) are included conjugated to the same fluorophore to be used as Tetraspanin Trio. This enables researchers to use two channels for their own biomarkers of interest if needed. We have built a modular kit where antibodies can be swapped for your own validated antibodies against targets of interest inside or outside EVs. These need to be conjugated to dSTORM-compatible dyes. 

You can find more information on fluorophores for dSTORM imaging here. The kit’s sample preparation protocol will guide you through the steps for incorporating your antibodies into the experiment and our image acquisition and analysis guide will provide you with the information needed when optimizing your antibodies.

Configuration example 488 channel 568 channel 647 channel
Kit components + antibody of choice Pan-EV stain-488 Tetraspanin Trio 568 own antibody 647
Kit components + cargo Add-on Pan-EV stain-488 Tetraspanin Trio 568 anti-Syntenin-1 647
Other tetraspanin Add-ons anti-CD9 488 anti-CD63 568 anti-CD81 647
Two antibodies of choice own antibody 488 Tetraspanin Trio 568 own antibody 647
Two antibodies of choice Pan-EV stain-488 own antibody 568 own antibody 647

EV analysis and quantification made simple

Purchase of the kit includes access to purpose built analysis tools within our CODI cloud platform, which allow seamless processing of single molecule localization data to obtain output summary data files.

  • Seamless upload data of acquired data to CODI cloud platform 
  • Visualization of individual EVs and their biomarkers with the nanoscale resolution
  • Analysis tools tailored to EV workflow:
    • Correction of spatial drift
    • Filtering of individual localizations
    • Cluster-based single molecule 
  • Extract quantitative data on EV populations based on raw data outputs (.csv format):
    • Biomarker positivity per sample
    • EV size distribution
    • Biomarker density per EV (localizations/cluster)
    • EV morphology parameters
  • Export high-resolution 3-color images from your EV samples 

ONI’s CODI cloud software uses clustering of single molecule data to enable study of EVs. Left image shows EVs captured using the EV Profiler Kit V2, which were imaged with the Nanoimager. Single molecule localization data is then clustered on CODI to help assess EV morphology features and determine levels of positivity for different biomarkers.

EV Profiler is powered by ONI’s Nanoimager and CODI platform

Combine the advantages of super-resolution imaging with powerful analysis of larger numbers of events. This allows detection and quantification of biomarker abundance with up to 3 biomarkers detected within the same sample using dSTORM, and reporting of EV morphology, including size and circularity. 

Whats in the box?

3 four lane flow-cell chips
6 lane stickers
2 stage securing magnets
Capture reagents
S4 capture (3 tubes)
Biotin capture CD9 (1 tube)
Biotin capture CD63 (1 tube)
Biotin capture CD81 (1 tube)
Detection probes
Pan-EV stain 488 (3 tubes)
Tetraspanin trio 568 (3 tubes, 3 antibodies mixed)
E1 standard EVs (1 tube)
S3, W1, F1, N1, P1, W2, C1 (3 tubes each)
BCubed imaging buffer parts A and B (3 tubes each)
Anti-Syntenin-1 647

Anti-CD9 568

Anti-CD63 568

Anti-CD81 568

Anti-CD9 488

Anti-CD81 647

Biotin anti-CD9

Biotin anti-CD63

Biotin anti-CD81


Request more information

Contact one of our sales specialists to request more information or to order EV Profiler V2

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